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RESEARCH

Dr. Yu-Wai Peter Lin

Dr. Teresa R. Petrino

Instructional Web Page

http://socrates.barry.edu/snhs-plin

RESEARCH

REPRODUCTIVE  ENDOCRINOLOGY

Developmental and Hormonal Aspects of

OOGENESIS

in Lower Vertebrates

KEY  WORDS

click on [**] to view references

Oocyte Maturation [**];   Germinal Vesical Breakdown (GVBD) [**];   Oogenesis;   Steroidogenesis [**];    Steroids; Cholesterol [**];  Progesterone;  17 Alpha,20 Beta-Dihydroxy-4-Pregnen-3-One [**]; Testosterone;   Estradiol [**]; Aromatase;   cAMP [**];   Protein Kinase C; Phorbol Ester [**];   Isoquinolinesulfonamide (H-8) [**]; Ovarian Follicles [**];   in vitro Culture [**]; Radioimmunoassay (RIA);   Bioassay [**];

Pituitary;  Gonadotropins [**](GTH) [**];    FSH;   LH; GTH I [**];   GTH II [**]; GTH Alpha Subunit;   GTH Beta Subunits [**][**]; Gonadopeptides;   Inhibin [**];   Activin; Endocrine;   Paracrine;   Autocrine [**]; Cloning;   DNA  Sequencing ;    Polymerase Chain Reaction (PCR); cDNA Library [**];   Nucleotide Sequences [**][**];

Polycomb Gene [**]; Rana pipiens (Leopard Frog) [**];  Xenopus laevis (African Claw Toad);  Fundulus heteroclitus (Mummichog;   Killifish) [**]; Danio rerio (Zebrafish) [**];  Amphibian [**];   Teleost [**]

Oocyte Maturation [**];  

Germinal Vesical Breakdown (GVBD) [**];

Oogenesis;   Steroidogenesis [**];    Steroids;

Cholesterol [**];  Progesterone; 

17 Alpha,20 Beta-Dihydroxy-4-Pregnen-3-One [**];

Testosterone;   Estradiol [**];

Aromatase;   cAMP [**];   Protein Kinase C;

Phorbol Ester [**];   Isoquinolinesulfonamide (H-8) [**];

Ovarian Follicles [**];   in vitro Culture [**];

Radioimmunoassay (RIA);   Bioassay [**];

Pituitary;

   Gonadotropins [**](GTH) [**];    FSH;   LH;

GTH I [**];   GTH II [**];

GTH Alpha Subunit;   GTH Beta Subunits [**][**];

Gonadopeptides;   Inhibin [**];   Activin;

Endocrine;   Paracrine;   Autocrine [**];

Cloning;   DNA  Sequencing ;    Polymerase Chain Reaction (PCR);

cDNA Library [**];   Nucleotide Sequences [**][**];

Polycomb Gene [**]

Rana pipiens (Leopard Frog) [**];   

Xenopus laevis (African Claw Toad);

Fundulus heteroclitus (Mummichog;   Killifish) [**];

Danio rerio (Zebrafish) [**];

Amphibian [**];   Teleost [**]

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NIH Funded Projects

National Institutes of Health

Grants NIH  Office  of  Extramural  Research   
http://grants.nih.gov/grants/oer.htm

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Molecular Dynamics of Gonadotropins

Abstract

Research Interest   and  Goals

To view other federally funded projects on Gonadotropins, 

Click on the CRISP logo below, and enter "Gonadotropins" as Search Terms in the Query Form and hit Submit Query

Computer Retrieval of Information
on Scientific Projects

http://crisp.cit.nih.gov/

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I have a long-standing interest in the endocrine and cellular mechanisms responsible for ovarian follicle development and oocyte maturation. 

This is a two prong project consisting of a molecular and a physiological component. 

The molecular portion focuses on the endocrine regulation of gonadotropins by sex-steroids and gonadal peptides at the pretranslational level. 

The physiological portion of the projects concentrates on the paracrine (local effect at neighboring cells)  and autocrine (local effect in the same cells) biological activities of the gonadal peptides in the ovary.

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Click on the following Links to view Citations about:

Fundulus heteroclitus

Rana pipiens

Zebrafish (Danio rerio)

Polycomb Gene

Permission to use the Copyright DNA image granted by:Paul A. Thiessen

http://cherubino.med.jhmi.edu/~paul

Fundulus  heteroclitus

copyright © 1998 by the Marine Biological Laboratory

________________________

Dept. of Natural Resources@Cornell.edu

http://www.sarep.cornell.edu/Sarep/fish/Cyprinodontidae/mummichog.html

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Fundulus  DNA    Blast   Citations

 

Fundulus Ovary  (40x)                Oocytes w/ Germinal Vesicle                              

 

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Fundulus heteroclitus (Classification)

 

References

 

 

Fundulus heteroclitus Gonadotropins

 

Lin, Y.-W. P., B. A. Rupnow, D. A. Price, R. M. Greenberg, and R. A. Wallace (1992). Fundulus heteroclitus gonadotropins 3. Molecular cloning and sequencing of beta subunits of two distinct gonadotropins (GTH I and GTH II) from pituitary cDNA library using the Polymerase Chain Reaction. Mol. Cell. Endocrinol. 85: 127-139.  Medline Accession No.  PMID: 1526312;   UI: 92405806.   /Abstract    

 

Links:   [Protein]  [DNA]  {Related Articles}   

 

Fundulus heteroclitus gonadotropin Nucleotide Sequences

GenBank Database http://www.ncbi.nlm.nih.gov/Web/Genbank/index.html

GenBank is the NIH genetic sequence database, an annotated collection of all publicly available DNA sequences ( Nucleic Acids Research 1998 Jan 1;26(1):1-7). There are approximately 2,570,000,000 bases in 3,525,000 sequence records as of April 1999. As an example, you may view the record for the neurofibromatosis gene. The complete release notes for the current version of GenBank are available. A new release is made every two months. GenBank is part of the International Nucleotide Sequence Database Collaboration, which is comprised of the DNA DataBank of Japan (DDBJ), the European Molecular Biology Laboratory (EMBL), and GenBank at NCBI. These three organizations exchange data on a daily basis.

Other Formats:
Links:

Fundulus heteroclitus gonadotropin II beta subunit (GTHII beta) mRNA, complete cds
GenBank gb|M87015|; FASTA Nucleotide FUNGTHII gi[213251] ; FASTA Protein gi|213252

Deduced  Fundulus GTH II beta  Protein Sequence:

>gi|213252|gb|AAB59963.1| gonadotropin II beta subunit MVCLFLGASSFIWSLAPAAAAFQLPRCQLLNQTISLEKRGCSGCHRVETTICSGYCATKDPNYKTSYNKA IQHVCTYGDLYYKTFEFPECVPGVDPVVTYPVALSCRCGGCAMATSDCTFESLQPDFCMNDIPFYH

Fundulus heteroclitus gonadotropin I beta subunit (GTHI beta) mRNA, complete cds
GenBank gb|M87014|; FASTA Nucleotide FUNGTHI gi[213249]; FASTA Protein gi|213250

Fundulus heteroclitus gonadotropin, alpha subunit (GTH) mRNA, complete cds.     GenBank gb|U12923|;FASTA Nucleotide gi|531886|; FASTA Protein gi|531887

Fundulus heteroclitus initiation factor (EIF-4A) mRNA, 3'end
GenBank gb|M89792|; FASTA Nucleotide FUNEIF4A gi [213247]; FASTA Protein gi|213248

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Blast (sequence similarity) Search : http://www.ncbi.nlm.nih.gov/BLAST/ 

Fundulus heteroclitus Gonadotropin   Sequences as Query

If you have any problems or questions with the results of this search
please refer to the BLAST FAQs

Distribution of 13 Blast Hits on the Query Sequence

Score E Sequences producing significant alignments: (bits) Value gb|M87015|FUNGTHII Fundulus heteroclitus gonadotropin II beta s... 888 0.0gb|L35096|MOZGONAIIB Morone saxatilis (striped bass) gonadotrop... 100 4e-19gb|L11722|AANGBS Acanthopagrus latus gonadotropin beta-subunit ... 76 6e-12gb|S51112|S51112 thyrotropin beta subunit [human, lymphocytes, ... 46 0.005dbj|D00578|RATTSHBA Rattus norvegicus mRNA for TSH beta subunit... 46 0.005

gb|L35096|MOZGONAIIB Morone saxatilis (striped bass) gonadotropin II beta-subunit mRNA, complete cds Length = 590 Score = 99.6 bits (50), Expect = 4e-19 Identities = 149/182 (81%), Positives = 149/182 (81%)

Query: 243 cagcatgtgtgcacttatggggatttgtactacaagacattcgagtttcctgaatgtgtg 302

  Sbjct: 259 cagcatgtgtgcacgtaccgggacttacactacaagacatttgagcttcctgactgtcca 318  

--------------------------------------

Query: 303 cccggagttgacccagttgtcacctaccccgttgccctgagctgccgctgtggaggctgt 362

  Sbjct: 319 cctggtgtggacccaaccgtcacctaccccgtggctcagagctgccactgcggccgctgt 378

--------------------------------------

Query: 363 gccatggccacatccgactgcaccttcgagagccttcagcctgacttttgcatgaatgac 422

  Sbjct: 379 gccatggacacgtctgactgtaccttcgagagcctgcagcccaacttctgcatgaatgac 438

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Fundulus heteroclitus Gonadotropin Subunits

GTH  II  beta

BLAST 1 - Fundulus heteroclitus gonadotropin II beta subunit (GTHII beta):  BlastN Search using gi|213251 as Query nucleotide sequence Blast1N

BLAST 2 - Fundulus heteroclitus gonadotropin II beta subunit (GTHII beta):  BlastP Search using gi|213252 as Query protein sequence Blast2P

GTH  I  beta

BLAST 3 - Fundulus heteroclitus gonadotropin I beta subunit (GTHI beta):  BlastN Search using gi|213249 as Query nucleotide sequence Blast3N

BLAST 4 - Fundulus heteroclitus gonadotropin I beta subunit (GTHI beta):  BlastP Search using gi|213250 as Query protein sequence Blast4P

GTH  alpha

BLAST 5 - Fundulus heteroclitus gonadotropin alpha subunit (GTH alpha):  BlastN Search using gi|531886 as Query nucleotide sequence Blast5N

BLAST 6 - Fundulus heteroclitus gonadotropin alpha subunit (GTH alpha):  BlastP Search using gi|531887 as Query protein sequence Blast6P

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List of gonadotropin sequences

Professor John Winfield, Univ.of Glasgow, UK

http://www.chem.gla.ac.uk/protein/glyco/swiss.html#Alpha

  ______________________________________

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Fundulus. Citations

http://www.mbl.edu/html/MRC/ABSTRACTS/Fundulus.2.cit.html

 

 

Steroidogenesis in Fundulus heteroclitus. I. Production of 17 alpha-hydroxy,20 beta-dihydroprogesterone, testosterone, and 17 beta-estradiol by prematurational follicles in vitro. Gen Comp Endocrinol 1989 Jan;73(1):147-56 PMID: 2920896, UI: 89153912

       

Fundulus heteroclitus gonadotropin(s). I. Homologous bioassay using oocyte maturation and steroid production by isolated ovarian follicles.
Gen Comp Endocrinol. 1987 Jul;67(1):126-41.
PMID: 3497837; UI: 87305481

Steroidogenesis in Fundulus heteroclitus V.: purification, characterization, and metabolism of 17 alpha,20 beta-dihydroxy-4-pregnen-3-one by intact follicles and its role in oocyte maturation.
Gen Comp Endocrinol. 1993 Oct;92(1):1-15.
PMID: 8262351; UI: 94085748

Steroidogenesis in Fundulus heteroclitus. IV. Dichotomous effects of a phorbol ester on ovarian steroid production and oocyte maturation.
J Exp Zool. 1992 Sep 1;263(3):254-64.
PMID: 1453155; UI: 93085336

Steroidogenesis in Fundulus heteroclitus. II. Production of 17 alpha-hydroxy-20 beta-dihydroprogesterone, testosterone, and 17 beta-estradiol by various components of the ovarian follicle.
Gen Comp Endocrinol. 1989 Nov;76(2):230-40.
PMID: 2591717; UI: 90076882

Effects of isoquinolinesulfonamide H-8 on Fundulus heteroclitus ovarian follicles: role of cyclic nucleotide-dependent protein kinases on steroidogenesis and oocyte maturation in vitro.
Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1997 May;117(1):75-81.
PMID: 9185330; UI: 97328815.

Oocyte maturation in the mummichog (Fundulus heteroclitus): effects of steroids on germinal vesicle breakdown of intact follicles in vitro.
Gen Comp Endocrinol. 1986 May;62(2):281-9.
PMID: 3781226; UI: 87055192

Analytical and experimental studies on the relationship between Na+, K+, and water uptake during volume increases associated with Fundulus oocyte maturation in vitro.
J Comp Physiol [B]. 1992;162(3):241-8.
PMID: 1613162; UI: 92308447

Oogenesis in Fundulus heteroclitus. I. Preliminary observations on oocyte maturation in vivo and in vitro.
Dev Biol. 1978 Feb;62(2):354-69. No abstract available.
PMID: 564305; UI: 78107517.

Oogenesis in Fundulus heteroclitus. III. Vitellogenesis.
J Exp Zool. 1983 Jun;226(3):441-57.
PMID: 6886667; UI: 83293294.

Major protein changes during vitellogenesis and maturation of Fundulus oocytes.
Dev Biol. 1985 Aug;110(2):492-8.
PMID: 4018410; UI: 85258612.

Serum hormone levels associated with spawning activity in the mummichog, Fundulus heteroclitus.
Gen Comp Endocrinol. 1988 May;70(2):345-54.
PMID: 3384313; UI: 88255815.

Pattern of vitellogenesis and follicle maturational competence during the ovarian follicular cycle of Fundulus heteroclitus.
Gen Comp Endocrinol. 1996 Jul;103(1):24-35.
PMID: 8812325; UI: 96411293.

Oocyte sensitivity to serotonergic regulation during the follicular cycle of the teleost Fundulus heteroclitus.
Biol Reprod. 1998 Jul;59(1):53-61.
PMID: 9674993; UI: 98337750

Functional heterologous gap junctions in Fundulus ovarian follicles maintain meiotic arrest and permit hydration during oocyte maturation.
Dev Biol. 1993 Nov;160(1):228-35.
PMID: 8224539; UI: 94040370.

Semilunar follicular cycle of an intertidal fish: the Fundulus model.
Biol Reprod. 1996 Apr;54(4):809-18.
PMID: 8924500; UI: 96249823.

 

Review Articles

A non-salmonid model for the study of fish reproduction. In: Proceedings of the Fourth International Symposium on Reproductive Physiology of Fish (A.P. Scott, J.P. Sumpter, D.E. Kime and M.S. Rolfe, eds.) pp.74-76.  Lin Y.-W. P., T.R. Petrino, and R.A. Wallace (1991).

Metabolic and developmental aspects of steroidogenesis in teleost ovaries. Compart.. Endocrinol. (Life Sci. Adv.) 8:33-45.  Lin Y.-W. P., T.R. Petrino, and R.A. Wallace (1989).

 

[See Related Articles] on Fundulus heteroclitus

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Rana  pipiens

northern leopard frog (photo by G.S. Casper)

© 1995, 1997 Milwaukee Public Museum, Inc.

http://www.mpm.edu/collect/vertzo/herp/atlas/Species/Rapip.html  

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Colorado Herpetological Society's Guide to the Reptiles and Amphibians of Colorado

http://www.coloherp.org/geo/species/sperapi.html

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Intrafollicular action of estrogen in regulating pituitary-induced ovarian progesterone synthesis and oocyte maturation in Rana pipiens: temporal relationship and locus of action. Gen Comp Endocrinol 1985 Jun;58(3):421-35 PMID: 3874115, UI: 85231984

In vitro estrogen modulation of pituitary and progesterone-induced oocyte maturation in Rana pipiens. J Exp Zool 1983 May;226(2):281-91 PMID: 6602863, UI: 83240493

Relative roles of theca and granulosa cells in ovarian follicular steroidogenesis in the amphibian, Rana nigromaculata.
Gen Comp Endocrinol. 1994 May;94(2):207-14.
PMID: 7926630; UI: 95011437.

Changes in the activities of steroidogenic enzymes during the development of ovarian follicles in Rana nigromaculata.
Gen Comp Endocrinol. 1993 Nov;92(2):225-32.
PMID: 8282172; UI: 94109657.

Evidence for follicle wall involvement in ovulation and progesterone production by frog (Rana pipiens) follicles in vitro.
Differentiation. 1982;22(2):79-84.
PMID: 6982185; UI: 83028280

Cholesterol mediation of progesterone production and oocyte maturation in cultured amphibian (Rana pipiens) ovarian follicles.
Biol Reprod. 1987 Jun;36(5):1219-28.
PMID: 3497671; UI: 87299916

Protein synthesis and steroidogenesis in amphibian (Rana pipiens) ovarian follicles: studies on the conversion of pregnenolone to progesterone.
Gen Comp Endocrinol. 1986 Sep;63(3):441-50.
PMID: 3493948; UI: 87163357

Role of follicle wall in meiosis reinitiation induced by insulin in Rana pipiens oocytes.
Am J Physiol. 1981 Jul;241(1):E51-6.
PMID: 7018265; UI: 81229865

Stimulatory and inhibitory effects of human follicular fluid on amphibian oocyte maturation and ovulation in vitro.
Differentiation. 1982;21(1):41-4.
PMID: 6978271; UI: 82165456.

Spontaneous maturation of follicular oocytes in Rana dybowskii in vitro: seasonal influences, progesterone production and involvement of cAMP.
J Exp Zool. 1989 Nov;252(2):190-9.
PMID: 2557374; UI: 90095381.

Role of protein kinase C activation in oocyte maturation and steroidogenesis in ovarian follicles of Rana pipiens: studies with phorbol 12-myristate 13-acetate.
Gamete Res. 1988 Nov;21(3):323-34.
PMID: 2854534; UI: 89232981.

Relation between acute pituitary responsiveness to gonadotropin releasing hormone (GnRH) and the ovarian cycle in the bullfrog, Rana catesbeiana.
Gen Comp Endocrinol. 1983 Jul;51(1):148-53.
PMID: 6411518; UI: 83288482.

Role of cAMP in modulating intrafollicular progesterone levels and oocyte maturation in amphibians (Rana pipiens).
Dev Biol. 1986 Oct;117(2):354-64.
PMID: 2428681; UI: 87005562.

Germinal vesicle migration and dissolution in Rana pipiens oocytes: effect of steroids and microtubule poisons.
Cell Differ. 1987 Apr;20(4):239-51.
PMID: 2884043; UI: 87215953.

Spontaneous oocyte maturation in Rana pipiens: Estrogen and follicle wall involvement. Gamete Research. 1985;12:11-28. Lin, Y.-W. P.  and A.W. Schuetz.

Studies on the mechanism of action of estradiol in regulating follicular progesterone levels: Effects on cAMP mediated events and 3beta-hydroxysteroid dehydrogenase. Devlop. Growth & Differ. 1988;30(6):611-618. Lin, Y.-W. P., H.B. Kwon, T.R. Petrino, and A.W. Schuetz.

Inhibitory action of the gonadopeptide Inhibin on amphibian (Rana pipiens) steroidogenesis and oocyte maturation. J Exp Zool. 1999;284(2):232-240. Lin, Y.-W. P., T. R. Petrino, A.M. Landin, S. Franco, and I. Simeus.  Abstract:PMID: 10404651, UI: 99332730

[See Related Articles] on Rana pipiens

on Inhibin and Oocyte Maturation

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Zebrafish  (Danio rerio)

Permission to use zebrafish image given by Pat Edwards:http://zfish.uoregon.edu/

The Fish Net Zebrafish Research Databases (Univ. Oregon)

SEARCH the Fish Net website

Zebrafish Book A guide for the laboratory use of zebrafish Danio* (Brachydanio) rerio

ZFIN - the on-line database of zebrafish information

ZEBRAFISH GENOMICS PROJECTS

FishScope, Confocal images and movies of early development from Mark Cooper at University of Washington

Embryonic and larval anatomy

Developmental , Zerba fish

Zebrafish Dev. Movie

ZEBRAFISH WEBSERVER (Zebrafish server at Harvard)

Report from the NIH Non-Mammalian Models Meeting, February 16, 1999 : Zebrafish Breakout Group Recommendations

Trans-NIH Zebrafish Coordinating Committee: Zebrafish Genomics Initiative

Zebrafish Earns Its Stripes in Genetic Screens-  Science 288:1160-1161, 2000

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Polycomb  Gene

Definition of Polycomb

Project in collaboration with Dr. Gerhild Packert

Danio rerio (zebrafish)

Putative Zebrafish Polycomb-like Gene

Sequence submitted to GenBank

Deduced from Clones B1;  B1.1; and B5 

Degenerate Oligonucleotide-Primed Polymerase Chain Reaction 

(DOP-PCR)

(Template: RT-PCR;   Primers: PolyF2, ZebraR6)

Primer Design ( K = T/G;  R = G/A;  N = A/T/G/C; )

                                            ( M = A/C;  Y = C/T )

        Polycomb F2 (Forward Primer):    5'      ATG GAG CTG CCN GCN GT      3'

                        Target  (Amino Acids)                    M   E   L   P   A   V       

        Zebra R6 (Reverse Primer):      3'       TG GTR GTC ATG GTC GGN K     5'

                      Sense Strand                 5'       AC CAY CAG TAC CAG CCN M  3'  

                    Target  (Amino Acids)           H        H        Q   Y   Q   P   

Polymerase Chain Reaction (PCR)

PCR condition   (Gel B1: Dec 4, 00;   Gel B1.1: Dec 7, 00)

Template:   Zebrafish cDNA (RNA from 24 hr embryos, RT-PCR)

Primers:   Degenerate Oligonucleotide Primers (internal),  Polycomb F2; Zebra R6

Enzyme:   Taq DNA polymerase (Promega)

Denature:    94 C,  30 sec;  Annealing:  55 C,  30 sec;  Extension:  72 C,  40 sec;   40 cycles

Expected PCR Product size:   around 400 bp

TA cloning (Promega)

DNA from Band B1 and Band 1.1 extracted and underwent TA cloning.

White colonies selected (Blue-white screening), Mini Plasmid Prep and DNA sequencing (University of Miami, DNA sequencing Core Lab)

Clone B1   (vector sequence removed using VacScreen)

>19_B1 Mar 30 01 20 to 420 bp  

(anti-sense strand sequence of Clone B1, Sequencing results using T7 as primer)

         Clone B1 (Sense vs Compliment of Antisense Sequence): Blast2 Result B1

           Clone B1.1 (Sense vs Compliment of Antisense Sequence): Blast2 Result B1.1

           Clone B5 (Sense vs Compliment of Antisense Sequence): Blast2 Result B5

        Pairwise Blasts (BLAST 2 Sequences) of the corresponding Sense Strand sequence (resulted from Sp6 sequencing) and the Compliment of Antisense Strand sequence (resulted form T7 sequencing) showed that there were 100% homology between the two sequencing (Sp6 vs. T7) results.   These would indicate to us that there were no sequencing error.

Clustal (Multiple Sequences) Alignment of Clone B1, B1.1 and B5:

ClustalW Result

Clones B1 and B5 are 100% identical.

Clone B1.1 shows three nucleotides [position 12* (T to A), 352* (C to A) and 402* (A to G)] different when compare to Clones B1 and B5.    

All three nucleotide changes did not result in a change of amino acid [ Proline (4)**, Arginine (118)** and Proline (134)**;  (silent mutation)].

** Amino Acid position;         * Nucleotide position

For detail, see ClustalW Alignment Result

NCBI blastx 

compares a nucleotide query sequence translated in all (6) reading frames against a protein sequence database

  Blastx  Search Results using the following as Query:  

B1-T7 -   Clone >19_B1 Mar 30 01 20 to 420 bp

B1.1-T7 -  Clone >20_B1.1 Mar 30 01 36 to 442 bp

B5-T7 -  Clone >24_B5 Mar 30 01 37 to 442 bp

B1-Sp6 -   Clone >B1-Sp6 Apr 06 01 64 to 466 bp

B1.1-Sp6 -  Clone >B1.1-Sp6 Apr 06 01 60 to 462 bp

B5-Sp6 -  Clone >B5-Sp6 Apr 06 01 60 to 462 bp

Example of Sequences producing significant alignments :

(Clone B1 as Query sequence)

ORF Finder (Open Reading Frame Finder)

ORF and Protein Sequence : B1-Sp6 Apr 06 01 64 to 466 bp

Clone B1: Zebrafish (Danio rerio) Polycomb mRNA partial cds

The Three (3) Reading Frames of Clone B1-Sp6 (Sense)

Zebrafish Polycomb-like Gene (partial) Sequence [Clone B1-Sp6 (Sense)]

submitted to GenBank Database (via BankIt) on April 6, 2001.

ACCESSION:  AY029300  [BankIt395137]

DEFINITION:   Danio rerio Polycomb mRNA partial cds.

AUTHORS:   Petrino,T.R., Lin, Y.-W.P., Packert,G.R., Nadal,C.N., Torrealday,S., and Greene,D.A.

GenBank Overview

http://www.ncbi.nlm.nih.gov/Genbank/GenbankOverview.html

Submissions to GenBank

Many journals require submission of sequence information to a database prior to publication so that an accession number may appear in the paper. NCBI has a WWW form, called BankIt, for convenient and quick submission of sequence data. Sequin, NCBI's stand-alone submission software for MAC, PC, and UNIX platforms, is also available by FTP. When using Sequin, the output files for direct submission should be sent to GenBank by electronic mail

OMIM™
Online Mendelian Inheritance in Man   

     http://www.ncbi.nlm.nih.gov/Omim/  

OMIM Definition of Polycomb:

http://www.ncbi.nlm.nih.gov/htbin-post/Omim/getmim

*603079 CHROMOBOX HOMOLOG 4, DROSOPHILA POLYCOMB CLASS; CBX4
*602770 CHROMOBOX HOMOLOG 2, DROSOPHILA POLYCOMB CLASS; CBX2
*602978 EARLY DEVELOPMENT REGULATOR 1; EDR1
*164831 LEUKEMIA VIRAL BMI-1 ONCOGENE, MOUSE, HOMOLOG OF; BMI1

POLYCOMB CLASS; CBX4

'Polycomb' (Pc) and other members of the Drosophila Polycomb group gene family are part of a cellular memory system that is responsible for the inheritance of gene activity by progeny cells. Pc shares a homologous domain known as the chromodomain with the Drosophila heterochromatin-binding protein, HP1. To isolate human Pc homologs, Satijn et al. (1997) screened a fetal brain cDNA library with a probe containing the nonchromodomain portion of a Xenopus Pc homolog (XPc). They recovered a cDNA encoding a predicted 558-amino acid protein that they designated human Pc2. Human Pc2 and XPc share 48% protein sequence identity. Northern blot analysis revealed that human Pc2 is expressed as a 2.8-kb transcript in all tissues tested with almost equal abundance. Using immunofluorescence, Satijn et al. (1997) found that human Pc2 colocalizes with another human Pc homolog, CBX2 (602770), in interphase nuclei, suggesting that the proteins are part of a complex. Based on the results of studies with mutant proteins and overexpression of wildtype protein, the authors suggested that human Pc2 is a repressor of protooncogene activity and that interference with human Pc2 function can lead to derepression of protooncogene transcription and subsequently to cellular transformation.

POLYCOMB CLASS; CBX2

'Polycomb' genes in Drosophila maintain the repressed state of homeotic and other developmentally regulated genes (Wedeen et al., 1986; Kuziora and McGinnis, 1988) by mediating changes in higher-order chromatin structure (Paro and Hogness, 1991). M33, a mouse homolog of polycomb, was isolated by means of the structural similarity of its chromodomain (Pearce et al., 1992). The fifth exon of M33 contains a region of homology shared by Drosophila and Xenopus. In Drosophila, its deletion results in the loss of polycomb function. Katoh-Fukui et al. (1998) disrupted M33 in mice by inserting a poly(A) targeting vector into its fifth exon. More than half of the resultant homozygous mutant mice died before weaning, and survivors showed male-to-female sex reversal. Formation of genital ridges was retarded in both XX and XY homozygous mutant embryos. Gonadal growth defects appeared near the time of expression of the Y-chromosome-specific Sry gene (480000), suggesting that M33 deficiency may cause sex reversal by interfering with steps upstream of Sry. Katoh-Fukui et al. (1998) pointed out that point mutations in the human SOX9 gene (114290) cause campomelic dysplasia (CMPD1) associated with male sex reversal. Because M33 is located distal to Sox9 on mouse chromosome 11 with a linkage of 5.3 cM, the possibility that Sox9 was affected coincidentally during gene targeting in ES cells was considered. However, this possibility was ruled out by other transgenic experiments. Katoh-Fukui et al. (1998) suggested that these homozygous M33-mutant mice represent the first case in which male-to-female gonadal sex reversal results from a defect in a known recessive gene. The human homolog of polycomb in Drosophila and M33 in mouse is presumably located on 17q since human SOX9 maps to 17q24.3-q25.1.   Gecz et al. (1995) isolated genomic clones of the human M33 homolog, which they designated CBX2. By analysis of somatic cell hybrids and by fluorescence in situ hybridization, Gecz et al. (1995) mapped the CBX2 gene to 17q25.

Pc2  Like Sequences

Homo sapiens Polycomb 2 homolog (hPc2) mRNA, complete cds. [GenBank Accession AF013956];  [FASTA Protein >gi|2317723];  [FASTA Nucleotide >gi|2317722]

Mus musculus transcriptional repressor Mpc2 mRNA, complete cds.   [GenBank Accession U63387];  [FASTA Protein >gi|2781392] ;  [FASTA Nucleotide >gi|2781391]

Gallus gallus mRNA for chromobox protein (CHCB3), complete cds.   [GenBank Accession AB005619];   [FASTA Protein >gi|3649787];  [FASTA Nucleotide >gi|3649786]

Xenopus laevis XPolycomb mRNA, complete cds. [xPc2] [GenBank Accession U39929];  [FASTA   Protein >gi|1086556]; [FASTA Nucleotide >gi|1086555]

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Polycomb 2 homolog [Homo sapiens]  (Pc2-like)  Related Sequences

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Pc1 Like Sequences

Drosophila (Pc), Xenopus (xPc1)  and Mouse (M33)

Drosophila melanogaster Pc gene for polycomb protein.  [GenBank Accession X55702];   [FASTA Protein >gi|603986];  [FASTA Nucleotide >gi|8321]

Xenopus laevis Polycomb homolog Pc1 (Pc1) mRNA, complete cds. [ xPc1] [GenBank Accession AF101438]; [FASTA   Protein >gi|3860185]; [FASTA Nucleotide >gi|3860184]

Mus musculus chromobox homolog 2 (Drosophila Pc class) (Cbx2), mRNA. [M33] [GenBank Accession NM_007623.1]; [FASTA   Protein >gi|6671692]; [FASTA Nucleotide >gi|6671691]

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Polycomb protein [Drosophila melanogaster] (Pc1-like)  Related Sequences

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Blastp: Blast search using Human Polycomb Protein (Pc2 like) as Query sequence.  BLASTP

Clustal : Multiple Aligment of Polycomb Proteins. ClustalW

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CLUSTAL W (1.74) multiple sequence alignment (Pc2 like)


gi|2317723       MELPAVG------------------------------------EHVFAVESIEKKRIRKGRVEYLVKWRGW 35
gi|2781392       MELPAVG------------------------------------EHVFAVESIEKKRIRKGRVEYLVKWRGW 35
gi|3649787       MELPAVG------------------------------------EHVFAVESIEKKRIRKGRVEYLVKWRGW 35
gi|1086556       MELPAVG------------------------------------EHVFAVESIEKKRIRKGRVEYLVKWRGW 35
gi|603986         MTGRGKGSKGKLGRDNATDDPVDLVYAAEKIIQKRVKKGVVEYRVKWKGW 50
              
gi|2317723       SPKYNTWEPEENILDPRLLIAFQNRERQEQLMGYRKRGPKPKPLVVQVPT 85
gi|2781392       SPKYNTWEPEENILDPRLLIAFQNRERQEQLMGYRKRGPKPNPLVVQVPT 85
gi|3649787       SPKYNTWEPEENILDPRLLIAFQNRERQEQLMGYRKRGPKPKPLVVQLPS 85
gi|1086556       SSKYNTWEPEENILDPRLLVAFQNRERQEQIMGYRKRGPKPKNNLVSMPS 85
gi|603986        NQRYNTWEPEVNILDRRLIDIYEQTNKSSGTPSKRGIKKKEKE--------PDPE 97
                                 * * * * * * *

            
gi|2317723       FARRSNVLTGLQDSSTDNRAKLDLGAQGKGQGHQYELNSKKHHQYQPHSK 135
gi|2781392       FARRSNVLTGLQDSSADNRAKLELGTQGKGQGHQYELNSKKHHQYQPHSK 135
gi|3649787       FARRSNILTGLQDPTVDTRPKLDLGSSGKSQQHQYELNSKKHHQYQPNGK 135
gi|1086556       FARRSNVLSGLHDSSGENRTKLDLGPIPKS-------HQYQLNSKKHHQYQPSGK 133
gi|603986         PESEEDEYTFTENDVDTHQATTSSATHDKE---SKKEKKHHHHHHHHHHIK 145
                                              

gi|2317723      TVTFKEYVTV 558
gi|2781392      TVTFKEYVTV 551
gi|3649787      TVTFKEYVTV 523
gi|1086556      TVTFKEYITV 521
gi|603986       ----------